P–522 Cervical secretion methylation profile is associated with the success of frozen-thawed embryo transfer - a proof-of-concept study

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y X Lee ◽  
C R Tzeng ◽  
Y M Hu ◽  
C H Chen ◽  
C W Chen ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Pregnancy Outcome ◽  
Embryo Transfer ◽  
Methylation Status ◽  
Endometrial Receptivity ◽  
Methylation Profile ◽  
Proof Of Concept ◽  
Gene Methylation ◽  
Non Invasive ◽  
Cervical Secretion

Abstract Study question Is cervical secretion gene methylation profile different between receptive and non-receptive endometrium and associated with implantation outcome in frozen-embryo transfer (FET) cycle? Summary answer The combination of candidate genes methylation profiles obtained from cervical secretion showed significant associations with pregnancy outcomes. What is known already Implantation failure remains a black box in reproductive medicine, and the exact mechanism of how endometrial receptivity is regulated is still unknown. Epigenetic modifications play a role in the gene expression pattern and may alter the endometrial receptivity in the human endometrium. Cervical secretion containing various implantation-related cytokines, and the gene methylation change can be used as a non-invasive molecular source that reflects the endometrium condition. Study design, size, duration In this retrospective case-control study, sixty-two women who entered the FET cycle (30 pregnant and 32 non-pregnant women) were enrolled. Participants/materials, setting, methods Cervical secretion was collected before embryo transfer from women enrolled in multicenter university-affiliated reproductive units. The DNA methylation status of six candidate genes was measured using quantitative methylation-specific PCR (qMSP). The correlation between methylation change and the pregnancy outcome was analyzed. Main results and the role of chance The candidate genes were selected from that associated with implantation with literature review and the original genome-wide DNA methylation data from NCBI GEO DataSets (GSE90060) which processed using bioinformatics analysis. Six candidate genes whose CpG-level methylation analysis with β-value statistically higher in receptive endometrium than in a pre-receptive endometrium were selected. All six candidate genes showed different degrees of correlation with the pregnancy outcomes. Among them, PRKAG2 methylation changes showed the highest correlation with the pregnancy outcome. A logistic regression model was used to evaluate the performance of a single gene or a combination of genes for implantation prediction. The results showed a statistically significant association between the methylation status of a combination of genes (PRKAG2, KRS1, HAND2) and the pregnancy outcome (p = 0.008), resulting in an optimal AUC of 0.7 (95% CI: 0.57 - 0.81) for implantation prediction. Limitations, reasons for caution The results obtained from a relatively small cohort size. A larger study and further comprehensive methylome investigations are warranted. Wider implications of the findings: This study is the first proof-of-concept study that cervical secretion methylation profile is associated with implantation outcome in a FET cycle, and showed potential as a non-invasive method for implantation prediction. Trial registration number non applicable

A Methylation Profile of Tumor Suppressor Genes Predicts a Poorer Survival in Patients with Refractory Anemia with Ringed Sideroblasts.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2780-2780
Author(s):  
Ana Valencia ◽  
Jose Cervera ◽  
Esperanza Such ◽  
Esther Gamero ◽  
Mariam Ibañez ◽  
...  
Keyword(s):  
Tumor Suppressor ◽  
Tumor Suppressor Genes ◽  
Methylation Status ◽  
Methylation Profile ◽  
Refractory Anemia ◽  
Aberrant Methylation ◽  
Gene Methylation ◽  
Suppressor Genes ◽  
Ring Sideroblasts ◽  
Aberrant Gene

Abstract Abstract 2780 Poster Board II-756 Patients with refractory anemia with ring sideroblasts (RARS) are considered to have good prognosis and anemia is usually managed with best supportive care and erythroid stimulating agents. Nowadays, no specific molecular marker related to outcome and disease progression has been identified. Several genes involved in cell cycle and apoptosis that may become inactivated by aberrant hypermethylation have been identified in patients with myelodysplastic syndromes (MDS) but the significance of a methylation profile (studying several genes at the same time) in RARS is unknown, mainly because the number of patients with RARS in published reports is rather low. To assess the implication of aberrant methylation in RARS, we studied the methylation status of 25 sequences of a set of tumor suppressor genes in 40 patients (median age 70 yr, 25 male gender) with RARS according to FAB criteria [WHO classification, RARS in 22 patients (55%); refractory citopenia with multilineage dysplasia and ring sideroblasts, 18 (45%)] by means of methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay. The MS-MLPA procedure (SALSA MLPA kit ME001, MRC-Holland, Amsterdam, The Netherlands) has been developed for detecting in a semi-quantitative manner changes in the methylation status of 25 tumor suppressor genes in a single experiment. Briefly, approximately 50 ng of DNA were denatured and hybridized with MLPA probes. Subsequently, the probes were ligated in half of every sample, whereas for the rest of the sample, ligation was combined with an endonuclease HhaI digestion resulting in ligation of the methylated sequences only. PCR was performed as described by the manufacturer, and then separated by capillary gel electrophoresis and quantified using the Genemapper software (ABI 310, Applied Biosystems, Foster City, CA). Quantification of the methylation status was done by dividing the peak area with the combined areas of the control probes lacking the target sequence of the HhaI. Finally, the relative peak area of each target probe from the digested sample was compared with those obtained from the undigested sample. Aberrant methylation was scored when the calculated methylation percentage was >10%. To validate the MS-MLPA method the methylation status of CDKN2B was also analyzed by methylation-specific PCR (MSP). Actuarial curves of OS were built by Kaplan-Meier method and differences between curves compared with log-rank tests. Small numbers precluded the use of multivariate methodology. The MS-MLPA analysis detected methylation of at least one gene in 17 patients (42.5%). The genes aberrantly methylated were CDKN2B (n = 8, 20%), RASSF1 (n = 7, 17%), RARB (n = 3, 7.5%), CDH13 (n = 3, 7.5%) and FHIT (n = 2; 5%). Of the 17 patients, five (12%) presented methylation in two genes (RASSF1-FHIT in 2, RASSF1-RARB in 1, RASSF1-CDH13 in 1, and CDKN2B-CDH13 in 1, who was the only patient who progressed to AML). The presence of aberrant gene methylation was not significantly associated with any clinical or biological characteristic or WHO morphological subtype. Patients with aberrant gene methylation had a significantly shorter overall survival (OS) than patients without methylated genes (median OS, 72 mo vs 114 mo, respectively; P = 0.03). Patients with hemoglobin level below 10 g/dL and platelet count below 150 ×109/L also had a significantly poorer OS (P= 0.01 and P= 0.02, respectively). As the majority of probes used in the MS-MPLA method detect methylation in only one CpG pair, the results of CDKN2B methylation were validated by MSP, which yielded the same methylation results than with MS-MPLA methodology. The 8 patients with methylated CDKN2B show a trend for a shorter survival than the remaining 32 with unmethylated CDKN2B (median 72 mo vs 114 mo, P = 0.08). The results of this study indicate that aberrant methylation of several tumor suppressor genes is observed in a substantial proportion of patients with RARS. As occurs in patients with high-risk MDS, our results suggest that aberrant gene methylation confers a poor prognosis in RARS, but these data and their potential independent value require confirmation in larger series that employ multivariate methods. Finally, these findings provide a strong rationale for the use of hypomethylating agents (e.g., azacitidine or decitabine) in patients with RARS. This work has been partially supported by ISCIII grants RD06/0020/0031 and CA08/00141. Disclosures: No relevant conflicts of interest to declare.

T2090 CAN1 Gene Methylation Profile in African Americans with Colon Cancer and Adenoma, New Candidate Genes

2008 ◽  
Vol 134 (4) ◽  
pp. A-617 ◽  
Author(s):  
Krishan Kumar ◽  
Mohammad Daremipouran ◽  
Pooneh Mokarram ◽  
Seyed Mehdi Nouraie ◽  
Edward L. Lee ◽  
...  
Keyword(s):  
Colon Cancer ◽  
African Americans ◽  
Candidate Genes ◽  
Methylation Profile ◽  
Gene Methylation

scholarly journals Physical Activity Level Influences MTHFR Gene Methylation Profile in Diabetic Patients

2021 ◽  
Vol 11 ◽  
Author(s):  
Tainá Gomes Diniz ◽  
Alexandre Sérgio Silva ◽  
Mayara Karla dos Santos Nunes ◽  
Mateus Duarte Ribeiro ◽  
João Modesto Filho ◽  
...  
Keyword(s):  
Physical Activity ◽  
Nutritional Status ◽  
Physical Activity Level ◽  
Methylation Status ◽  
Methylation Profile ◽  
Activity Level ◽  
Active Group ◽  
Diabetic Patients ◽  
Gene Methylation ◽  
Partially Methylated

IntroductionMTHFR methylation status is associated with microvascular complications in diabetes, but the factors influencing this profile remain unknown.ObjectiveThe aim of this study was to evaluate the influence of physical activity level and nutritional status on the methylation profile of the MTHFR gene in patients with type 2 diabetes mellitus (T2DM).MethodsA total of 111 patients, 43 men and 68 women diagnosed with DM (7.0 ± 2.3 years), answered the International Physical Activity Questionnaire (IPAQ) and underwent blood collection for biochemical analysis, DNA extraction, and MTHFR gene methylation profile determination.ResultThe comparison of the methylation pattern showed that the partially methylated profile predominates in the insufficiently active group (85%), which does not occur in the sufficiently active group (54%) (p = 0.012). No differences were found in the nutritional status comparison. Logistic regression including overweight, waist circumference, gender, age, time of DM, hypertension, dyslipidemia, smoking, alcoholism, and family DM revealed that the association of the level of physical activity with methylation profile proved to be independent of these confounding variables. Considering the partially methylated profile as a result, being physically inactive favors the partially methylated MTHFR pattern in patients with DM.ConclusionWe concluded that insufficient physical activity is associated with partially methylated pattern of MTHFR promoter.

New candidate genes to predict pregnancy outcome in single embryo transfer cycles when using cumulus cell gene expression

2012 ◽  
Vol 98 (2) ◽  
pp. 432-439.e4 ◽  
Author(s):  
Sandra Wathlet ◽  
Tom Adriaenssens ◽  
Ingrid Segers ◽  
Greta Verheyen ◽  
Ronny Janssens ◽  
...  
Keyword(s):  
Gene Expression ◽  
Candidate Genes ◽  
Pregnancy Outcome ◽  
Embryo Transfer ◽  
Cumulus Cell ◽  
Single Embryo Transfer ◽  
Cell Gene Expression ◽  
Cell Gene

scholarly journals Gene Expression Profiles Associated with Radio-Responsiveness in Locally Advanced Rectal Cancer

Biology ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 500
Author(s):  
Jeeyong Lee ◽  
Junhye Kwon ◽  
DaYeon Kim ◽  
Misun Park ◽  
KwangSeok Kim ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Expression Profiles ◽  
Cell Cycle Control ◽  
Methylation Status ◽  
Locally Advanced ◽  
Gene Expression Profiles ◽  
Advanced Rectal Cancer ◽  
Bioinformatic Analysis ◽  
Locally Advanced Rectal Cancer ◽  
Genes Encoding

LARC patients were sorted according to their radio-responsiveness and patient-derived organoids were established from the respective cancer tissues. Expression profiles for each group were obtained using RNA-seq. Biological and bioinformatic analysis approaches were used in deciphering genes and pathways that participate in the radio-resistance of LARC. Thirty candidate genes encoding proteins involved in radio-responsiveness–related pathways, including the immune system, DNA repair and cell-cycle control, were identified. Interestingly, one of the candidate genes, cathepsin E (CTSE), exhibited differential methylation at the promoter region that was inversely correlated with the radio-resistance of patient-derived organoids, suggesting that methylation status could contribute to radio-responsiveness. On the basis of these results, we plan to pursue development of a gene chip for diagnosing the radio-responsiveness of LARC patients, with the hope that our efforts will ultimately improve the prognosis of LARC patients.

Sign in / Sign up

Export Citation Format

Share Document